Preparation, process and a regenerative method and technique for prevention, treatment and glycemic control of diabetes mellitus

ABSTRACT

A medicinal preparation, a process, a nutritional composition and regenerative and insulin potentiating method for humans and also for mammals for prevention, treatment and management/glycemic control of diabetes mellitus by obtaining optimum glucose level in blood, by administering an extract of  Costus pictus  to produce above effect in a dose of 500-2000 mg/day in humans and 50-200 mg/kg/day in rats. A method for treating hyperglycemia in a patient by administering a medicinal preparation of  Costus pictus  D. Don. A method for long term management of diabetes by avoiding the problems associated with tight control of blood glucose concentrations, i.e., hypoglycemia tolerance and seizures, while simultaneously avoiding the problems associated with conventional moderate control of blood glucose concentrations, i.e., pathological complications associated with hyperglycemia, such as nephropathy, retinopathy, etc. The medicinal preparation further affords an insulin potentiating action in addition to decreasing blood glucose and/or glycosylated hemoglobin levels.

RELATED APPLICATIONS

This application is a divisional of co-pending U.S. application Ser. No.13/064,565, Mar. 31, 2011, which is a divisional of U.S. applicationSer. No. 12/591,067, filed Nov. 6, 2009, which is a divisional of U.S.application Ser. No. 12/081,849, filed Apr. 22, 2008 issued as U.S. Pat.No. 7,635,495, which is a divisional of U.S. application Ser. No.11/808,381, filed Jun. 8, 2007, which issued as U.S. Pat. No. 7,378,113,which is a divisional of U.S. application Ser. No. 11/418,250, filed May5, 2006, which issued as U.S. Pat. No. 7,255,886 on Aug. 14, 2007, whichis a continuation of PCT Application Serial No. PCT/IN2004/000019, filedJan. 28, 2004. The entire disclosures of the prior applications U.S.application Ser. Nos. 12/591,067, 12/808,381 and 11/418,250 andInternational Application No. PCT/IN2004/00019 are hereby incorporatedby reference.

FIELD

This disclosure relates to an innovative and path-breaking techniquethat treats Diabetes mellitus and to obtain optimum glycemic control intype I and type II diabetic patients. The disclosure also provides aneco-friendly method and manner of extracting the plant Costus pictus D.Don.

Syn: Costus mexicanus.

This disclosure provides a method of treating Diabetes mellitus toobtain optimum glycemic level in blood.

BACKGROUND References

The following papers provide useful background information, for whichthey are incorporated herein by reference in their entirety.

-   1) U.S. Pat. No. 6,632,461, issued Oct. 14, 2003 “Use of tropical    root crops in effective intervention strategies for treating    difficult and complex cases and chronic diseases” by Slimak.-   2) U.S. Pat. Application 20040002423 A1, Jan. 1, 2004, “Remedies” by    Ohnogi, Hiromu.-   3) Zimmet P, Alberti K G, Shaw J. Global and societal implications    of the diabetes epidemic. Nature 2001; 414: 782-787.-   4) King Aubert R E, Herman W H. Global burden of Diabetes,    1995-2025: prevalence, numerical estimates, and projections.    Diabetes Care 1998; 21: 1414-1431.-   5) Ramachandran A, Snehalatha C, Latha E, Vijay V, Viswanathan M.    Rising prevalence of NIDDM in an urban population in India.    Diabetologia 1997; 40: 232-237.-   6) Harris M I, Fleggal K, Cowie C, Eberhardt M S, Goldstein D E,    Little R R, Wiedmeyer H M, Byrd-Holt D D. Prevalence of diabetes,    impaired fasting glucose and impaired glucose tolerance in U.S    adults. The Third National Health and Nutrition Examination Survey,    1988-1994. Diabetes Care 1998; 21: 518-524.-   7) Type 2 Diabetes in children and adolescents. American Diabetes    Association. Diabetes Care 2000; 23: 381-389.-   8) Katzung G. Basic and Clinical Pharmacology, International    Edition, (8th Edition) edited by Bertraram, Pages 2001, 723-730-   9) National Health Interview Survey and (1999-2001) National Health    and National Examination Survey estimates projected to year: 2002.-   10) Grifin B. W. Prevention of diabetic cataract and neuropathy in    rats by two new aldose inhibitors. Invest Ophthal. Vis. Sci. (1984)    25:136.-   11) Rao R M, Salem F A, Gleason-Jordan I. Antidiabetic effects of    dietary supplement “Pancreas Tonic” J. Natl. Med. Assoc., (1998)    90:614-618.-   12) Asatoor A M & King E J, Simplified colorimetric blood sugar    method, Process Biochem. 1954 Jan. 16; 56 (325^(th) meeting).-   13) Zhou, J. et al., Diabetes, 1999; 48 (12): 2358-2366.-   14) Xu, G. et al., Diabetes, 1999; 48 (12):2270-2276.

BACKGROUND

In this section, we discuss several aspects of related work, includingbackground and conventional technologies.

Globally, the number of people with diabetes is expected to rise fromthe current estimate of 150 millions to 220 millions in 2010 and 300millions in 2025. The prevalence is increasing in the developingcountries such as India, particularly in urban areas. The estimatednumber of diabetes patients in India was 19.4 million in 1995 and areexpected to be 57.2 million in 2025 (W.H.O). In the United States, it isestimated that as of 2002, 18.2 million people (6.3% of the totalpopulation) were diabetic. About 2.06 lakhs of people under 20 years ofage have diabetes (0.25% of all people in this age group). Approximatelyone in every 400-500 children and adolescents has Type-I diabetes. Inthe age group of 20 years or older, 18 million (8.7% in men and 9.3% inwomen) have diabetes. In people above the age of 60 years, 8.6 million(18.3% of all people in this age group) have diabetes.

There are mainly two types of diabetes. Type-I diabetes which waspreviously called insulin dependent diabetes mellitus (IDDM) orjuvenile-onset diabetes. This develops when the body's immune systemdestroys pancreatic beta cells (β-cells), which are the only cells inthe body that make the hormone insulin that regulates blood glucoselevel. The pancreas comprises two glandular tissues, one, is acollection of cells that form the exocrine function of the pancreaswhere these exocrine cells synthesize and release digestive enzymes intothe intestine; the second tissue comprises the endocrine function of thepancreas which synthesize and release hormones into the circulation. Ofprime importance for the endocrine function of the pancreas are the betacells. These cells synthesize and secrete the hormone insulin. Thehormone insulin plays a vital role in maintaining normal physiologicalglycemic levels. There are molecules that are effectors of the endocrinecells of the pancreas. Glucoincretins are an example of such molecules.Incretins potentiate glucose-induced insulin secretion from thepancreas. In Type-I diabetes there is decreased insulin production andthe circulating insulin level is very low. Type-I diabetes usuallystrikes children and young adults, although the disease onset can occurat any age. It accounts for 5-10% of all diagnosed cases of diabetes.Risk factors for Type-I diabetes may include autoimmune, genetic, andenvironmental factors.

Type-II diabetes was previously called non-insulin dependent diabetesmellitus (NIDDM) or adult onset diabetes. It usually begins as insulinresistance, a disorder in which the cells of the body fails to respondto insulin properly. As the need for insulin rises, the pancreasgradually loses its ability to produce insulin. Beta cells of pancreaticislets are dysfunctional. Type-II diabetes is associated with older agegroup, obesity, family history of diabetes, history of gestationaldiabetes, impaired glucose metabolism, physical inactivity, andrace/ethnicity. In recent years, Type-II diabetes is increasingly beingdiagnosed in children and adolescents. Type II diabetes is thepredominant form of diabetes world wide, accounting for 90% of casesglobally. An epidemic of Type II diabetic patients is underway in bothdeveloped and developing countries, although the brunt of the disorderis felt disproportionally in non-European populations, especially inIndia.

Type II diabetes has become one of the world's most important publichealth problems. It is currently thought to occur in geneticallypredisposed individuals, who are exposed to a series of environmentalinfluences that precipitate the onset of clinical disease. Sex, age andethnic background are important factors in determining the risk for thedevelopment of Type II diabetes. The disorder is more common in thefemales.

Age is also an important factor. Type II diabetes has been viewed in thepast as a disorder of aging with an increasing prevalence with age. Thisremains true today. However, a disturbing trend has become apparent inwhich the prevalence of obesity and type II diabetes in children isrising dramatically. In the past, it was believed that the overwhelmingmajority of children with diabetes had type I diabetes, with only 1-2%children considered to have type II or other rare forms of diabetes.Recent reports suggest that as many as 8-45% of children with newlydiagnosed diabetes have non-immune-mediated-diabetes. The majority ofthese children have type II diabetes, but other types are also beingidentified. Normal insulin secretory function is essential for themaintenance of normal glucose tolerance, and abnormal insulin secretionis invariably present in patients with type II diabetes.

Gestational diabetes is a form of glucose intolerance that is diagnosedin some women during pregnancy. It is more common among obese women andwomen with a family history of diabetes. During pregnancy, gestationaldiabetes requires treatment to normalize maternal glucose level in orderto avoid complications in the infant. After pregnancy, 5-10% of womenwith gestational diabetes are found to have type-II diabetes. Women whohave had gestational diabetes have a 20% to 50% chance of developingdiabetes in the next 5-10 years.

Other specific types-of diabetes result from specific genetic conditions(such as maturity onset diabetes of youth), surgery, drug-induced,malnutrition, infections, and other illnesses. Such types account for1-5% of all diagnosed cases of diabetes.

Type-I diabetic patients have virtually no functional (3-cells, canneither respond to variations in circulating glucose nor maintain abasal secretion of insulin, and the treatment principle is to achieveblood glucose concentration as close to normal as possible to reduce therisk of diabetic complications. Previous proposals for treatment includeapplication of exogenous bioactive Glucagon-like peptide-1 (7-36) amide(“GLP-1”) or its analogs, to either stimulate islet cell regeneration invivo, or to obtain pancreatic cells from diabetes mellitus patients andto treat such cells ex vivo in tissue culture using bioactive GLP-1.This ex vivo treatment was considered to facilitate regeneration and/ordifferentiation of Islet cells which could then synthesis and secreteinsulin or glucagon.

However, such a treatment regime requires the enteral or parenteralapplication of bioactive GLP-1 to patients, including the possibility ofsurgery. It is one aspect to obviate the need for surgical treatment,enteral or parenteral applications of bioactive GLP-1.

There are two types of treatments depending on the patient's condition.Intensive therapy—with insulin administration by insulin pump or 3-4 ormore daily injections with a goal to achieve blood glucose levelsbetween 70-120 mg/dl before meals, less than 180 mg/dl after meals andweekly 3 AM measurement greater than 65 mg/dl and HbA1C (glycosylatedhaemoglobin) value within the normal range (6.05% or less).

Conventional therapy consists of one or two daily injection of insulinincluding mixed intermediate and rapid acting insulins daily, with selfmonitoring of urine and blood glucose. Intensive therapy gives goodglycemic control and decreases the risk of retinopathy by 47%,microalbuminuria by 34%, while secondary intervention causes 43%protection.

According to the American Diabetes Association's revised guidelines for2002, the goal of therapy is to achieve average prepandial plasmaglucose concentration in the range of 90-130 mg/dl, average bedtimeplasma glucose values between 110-150 mg/dl and HbA1C values less than7%.

In Type II diabetic patients, beta cells (β-cells) of the Islets ofLangerhans are intact, but dysfunctional. They are not secreting enoughinsulin needed, there is also insulin resistance and the cells do notrespond to insulin. The major predisposing factor is obesity. Hencetreatment is initiated with dietary changes and exercise and if thepatient does not improve, drugs are prescribed. The revolution in thetreatment of Type II diabetes since 1995 has been the release ofmultiple new classes of drugs that independently address differentpathophysiological mechanisms that contribute to the development ofdiabetes.

The available oral anti-diabetic drugs can be divided by their mechanismof action as shown in Table 1.

TABLE 1 Orally administered anti-diabetic drugs. Approved MechanismClass Drugs Of action Limitations Sulphonylureas 2 (2^(nd) gen) Act onpancreatic Insulin (Insulin secretagogues) 4 (1^(st) gen) β Cells torelease resistance insulin Biguanides only one decrease glucose lacticacidosis (Insulin sensitizers) (Metformin) release by liver improvesinsulin sensitivity diminish insulin fluid retention weight gainThiazolidinediones only one resistance & hypoglycemia, (Insulinsensitizers) (Glitazones) improve insulin cannot be used sensitivity inpresence of heart & liver disease ∝Glucosidase 2 (Acarbose, slow theflatulence, inhibitors Miglitol) absorption of diarrhoea, carbohydratesabdominal pain

Even though a variety of modern drugs are available for the treatment ofdiabetes mellitus, as in above table none of them are without sideeffects. Insulin even though very potent, remains the drug of choice forType-I diabetics and also for type-II diabetics who do not obtainglycemic control with oral anti-diabetic drugs. The main disadvantage ofinsulin is that it is not orally effective because it is destroyed uponoral administration, and hence has to be given by injection.

BACKGROUND ART

Diabetes mellitus was known to ancient Indians as early as sixth centuryB.C. Charaka in his “Charaka Samhita” has mentioned the sweetness ofurine in addition to polyuria. The Indian physician Susruta in 500 A.Ddescribed the disease as “Madhumeha” meaning rain of honey, (due to thephenomenon of attracting ants near the urine of a diabetic patient) withsymptoms of foul breath, voracious appetite and langour. Other earlyIndian writings like Astanga-hrayda, Bhava-prakasa, Madhav-nidana etc.have also described diabetes.

In ancient traditional medicine, the treatment of diabetes was attemptedwith different indigenous plants or their combinations administeredorally. These anti-diabetic agents can be divided into two groupsaccording to their source of origin: vegetable or herbal preparationsand mineral preparations. Vegetable or herbal agents administered as“asawas” and “arishtas’ constitute the majority of Indian indigenousanti-diabetic drugs. Mineral preparations are given as “Bhasmas” arecomparatively fewer. The main stay of treatment was dietary therapy.

The high costs of modern treatment of diabetes indicate a great need forthe development of alternative strategies for the prevention andtreatment of diabetes. Approximately 90% of the population in ruralareas of developing countries still relies on traditional medicines fortheir primary health care.

Since antiquity, diabetes has been treated with plant medicines. Morethan 1200 species of plants have been screened for anti-diabeticactivity on the basis of Ethanopharmacology or on random basis. Theseproducts range from marine algae and fungi to phylogenetically advancedclasses of plants. There are about 200 pure compounds from plant sourceswith reported blood sugar lowering effect. Recent scientificinvestigations on animals and humans had confirmed the efficacy of manyof these preparations, some of which are very effective, relativelynon-toxic.

They are broadly divided into (a) those used as part of diet (b) thoseused as herbal drugs. Onion (Allium cepa), Garlic (Allium salivum),Fenugreek—(Trigonella foenum-graecum), Momordica charantia (bittergourd/Kaippaka), Coccinia grandis (Linn), Cinnamomum tamela (Indian bayleaf), Muraya koengei leaves and Cyamopsis teiragonoloba (Cluster beans)are vegetables which form a part diabetic diet. Emblica officinalis(Indian goose berry, amla), Curcuma longa (Turmeric), Gymnema sylvestre(Chakkara kolli), Pterocarpus marsupium (Venga) etc are some of herbalplants, found to have antidiabetic properties.

Many more herbs exhibit anti-diabetic action, but they have not beenstudied systematically. Unlike what was happening a few decades ago,anti-diabetic herbal formulations are marketed now in plenty in the formof powders, tablets, capsule and liquid preparations. Most of these havenot undergone controlled clinical trials. Usually herbs with reputedanti-diabetic activity as documented in standard ayurvedic books orknowledge gained from experienced ayurvedic practitioners are taken upand mixtures are prepared with arbitrary dosage. Since most of these arenon-toxic it is not causing much harm. It is true that some of thesepreparations are effective.

Hypoglycemic actions of some of the herbs with reputed use in thetraditional system of medicine, Ayurveda, and used by the localAyurvedic practitioners were reviewed with particular reference to thefamily ‘Zingiberaceae.’ Curcuma longa has been used in Indian andChinese systems of medicine for a long time. The dried rhizome, commonlyknown as turmeric, is used for culinary purposes and is incorporatedinto many ayurvedic preparations. Turmeric mixed with amla (Emblicaofficinalis) fruit powder is used in the form of tablets for thetreatment of Diabetes. Ginger, which belongs to the familyZingiberaceae, is also a rhizome. The fresh rhizome is used as a spiceand in the preparations of curries. Ginger is also used as a driedpowder for many Ayurvedic preparations, including those for thetreatment of diabetes.

Interacting defects in muscle, liver, adipose tissue and pancreas,generate the pathogenic milieu that results in diabetes. Various classesof oral antihyperglycemic agents are currently available that target thedifferent pathologic factors contributing to diabetes, for example,α-glucosidase inhibitors that delay intestinal carbohydrate absorption,biguanidines that target hepatic insulin resistance, insulinsecretagogues that increase pancreatic insulin secretion,thiozolidonediones as insulin sensitizers to target adipocyte and muscleinsulin resistance, and intestinal lipase inhibitors to inhibit fatabsorption and promote weight loss in obese patients. A large number ofplant-derived medications have also been described to effect glycemiccontrol. However, none of these address the various aspects of diabetesconcurrently, namely, control of blood sugar, regeneration of the betacells of pancreas resulting in increased secretion of insulin, reducinginsulin resistance, and correcting the decreased hepatic glycogensynthesis.

For this study, I selected an ornamental plant of the family Costaceaeknown as Costus pictus D. Don, which formerly belonged to the familyZingiberaceae and resembles ginger and turmeric plants but for the factthat the above said ornamental plant is not being used in any of theoriental treatment regimen for medicinal purpose, particularly fordiabetes.

I hypothesized that since plants of the family Zingiberaceae, liketurmeric are propagated as anti-diabetics, some other plants of the samefamily Zingiberaceae might be effective in the management of Type I andType II diabetes.

SUMMARY

The disclosed teachings provide an extract of Costus pictus D. Don. Thedisclosure provides a preparation containing an extract of Costus pictusD. Don selected from a group consisting of a medicinal preparation, anutritional composition, and combinations thereof.

The disclosure provides a pharmaceutical composition of an extract ofCostus pictus D. Don. The disclosure provides a dosage form or anextract of Costus pictus D. Don. The disclosure provides a dosage formof an extract of Costus pictus D. Don for oral administration. Dosageforms of the extract are selected from the group consisting of acapsule, tablet, granule, sachet, powder, paste, ointment, infusion,injection, ampoule, solution, suspension, emulsion, pill, sustainedrelease formulation and combinations thereof.

The disclosure provides a method of treatment of diabetes mellitus byadministering an extract of Costus pictus D. Don to a mammal. Mammalsinclude streptozotocin induced diabetic rats and humans. The method oftreatment of diabetes mellitus includes administering a dose of about 50mg/kg/day to about 200 mg/kg/day of an extract of Costus pictus D. Donto the streptozotocin induced diabetic rat. The method of treatment ofdiabetes mellitus includes administering a dose of about 500 mg/day toabout 2000 mg/day of an extract of Costus pictus D. Don to a human inneed of treatment, prevention or management of diabetes.

The disclosure provides a method of management of diabetes in a mammalby administering an extract of Costus pictus D. Don.

The disclosure provides a method of treatment, prevention or managementof diabetes in a mammal by administering an extract of Costus pictus D.Don.

The disclosure provides a method of inducing hypoglycemia in a mammalfor treatment, prevention or management of diabetes in the mammal byadministering an extract, of Costus pictus D. Don to the mammal.

The disclosure provides a method for regenerating beta cells ofpancreatic islets in a mammal for treatment, prevention or management ofdiabetes in the mammal by administering an extract of Costus pictus D.Don to the mammal.

The disclosure provides a method for increasing the capacity of insulinproducing cells in a mammal for the treatment, prevention or managementof diabetes in the mammal by administering an extract of Costus pictusD. Don, whereby the endogeneous insulin producing cells of the mammalbecome more effective insulin producers.

The disclosure provides a method for increasing the capacity of insulinproducing cells in a mammal for the treatment, prevention or managementof diabetes in the mammal by administering an extract of Costus pictusD. Don, whereby damaged beta cells present in the pancreas regenerateinto insulin producing cells.

The disclosure provides a method of controlling normal liver glycogen ina mammal for the treatment, prevention or management of diabetes in themammal by administering an extract of Costus pictus D. Don.

The disclosure provides a method of preparing an extract of Costuspictus D. Don including:

collecting a starting material comprising Costus pictus D. Don; and

treating the starting material with low molecular weight alcohols, lowmolecular weight alcohols mixed with water, halogenated hydrocarbons,organic ethers, low molecular weight esters, other organic solvents, lowmolecular weight ketones, water and combinations thereof to prepare theextract, wherein the starting material comprises a fresh whole plant ofCostus pictus D. Don, fresh leaves of Costus pictus D. Don, dried leavesof Costus pictus D. Don, or combinations thereof.

The disclosure provides a method of preparing an extract of Costuspictus D. Don including:

-   -   collecting a starting material comprising Costus pious D. Don;

refluxing the starting material with a first low molecular weightalcohol to prepare a mixture I;

filtering the mixture I to prepare a filtrate I and a residue I;

concentrating the filtrate I to prepare a concentrate I; and

drying the concentrate I to prepare a fraction I,

wherein the starting material comprises a fresh whole plant of Costuspictus D. Don, fresh leaves of Costus pictus D. Don, dried leaves ofCostus pictus D. Don, or combinations thereof, and wherein the fractionI is designated as the extract.

The disclosure provides a method of preparing an extract includescollecting fresh leaves of Costus pictus D. Don, cleaning the freshleaves to prepare cleaned leaves, drying the cleaned leaves to preparedried leaves, powdering the dried leaves to prepare a dried powder, andrefluxing the dried powder with a first low molecular weight alcohol toprepare the extract.

BRIEF DESCRIPTION OF THE DRAWINGS

The above objectives and advantages of the disclosed teachings willbecome more apparent by describing in detail preferred embodimentsthereof with reference to the attached drawings in which:

FIG. 1 shows a procedure to prepare one or more extracts from freshleaves of Costus pictus D. Don.

FIG. 2 provides a graph showing blood glucose level in untreated normalrats (control); normal rats treated with various fractions, namely,Fraction I (fr I), Fraction III (fr III), Fraction IV (fr IV) andFraction V (fr V); normal rats treated with glybenclamide and metformin.The x-axis provides duration in hrs after feeding extract to fastingnormal rats. The y-axis provides blood glucose concentration in mg/dl.

FIG. 3 provides a graph showing glucose tolerance test in normal rats,which include untreated (control) rats, normal rats treated with 50mg/kg (amount of extract/body weight of rat), normal rats treated with100 mg/kg, normal rats treated with 200 mg/kg and normal rats treatedwith 1 gm/kg. The x-axis provides duration in hrs, wherein the zero hrsrefers to the fasting blood sugar level (FBS). The y-axis provides bloodsugar level in mg/dl.

FIG. 4 provides a graph showing glucose tolerance test in diabetic rats.Group A were untreated normal rats (control), Group B were rats treatedwith streptozotocin to induce diabetes followed by treatment withextract and Group C refers were rats treated with streptozotocin but notwith extract. The x-axis provides duration in hours over which the bloodglucose concentration was monitored. The y-axis shows blood glucoseconcentration in mg/dl. The arrow indicates the time of glucose loadingin the rats.

FIG. 5 provides a graph showing effect of extract administration onblood glucose level. Group A were untreated normal rats (control), GroupB were rats treated with streptozotocin to induce diabetes followed bytreatment with extract, and, Group C refers were rats treated withstreptozotocin but not with extract. The x-axis shows the duration indays over which the glucose level was monitored. The y-axis shows bloodglucose concentration in mg/dl.

FIG. 6 provides a graph showing effect of extract administration onlevels of liver glycogen. Group A were untreated normal rats (control),Group B were rats treated with streptozotocin to induce diabetesfollowed by treatment with extract and Group C refers were rats treatedwith streptozotocin but not with extract. The x-axis provides thedifferent groups. The y-axis provides glycogen level in mg/gm.

FIG. 7 provides a graph showing the effect of extract administration onbody weight of rats. Group A were untreated normal rats (control), GroupB were rats treated with streptozotocin to induce diabetes followed bytreatment with extract and Group C refers were rats treated withstreptozotocin but not with extract. The x-axis provides the timeinterval over which body weight was monitored. The y-axis provides bodyweight in gm.

FIG. 8 provides a graph showing the effect of extract administration onblood glucose level in a normal and in a diabetic human. The x-axisprovides the time interval over which blood glucose was monitored. They-axis provides glucose concentration in mg/dl.

DETAILED DESCRIPTION

The present invention relates to the use of a new plant extract in thetreatment of diabetes mellitus. Diabetes mellitus is a group ofmetabolic disorders characterized by chronic hyperglycemia due torelative insulin deficiency, resistance or both. There is defect ininsulin production, insulin action or both. The disclosure provides aninnovative method and manner of preparing a unique medicinal preparationfrom Costus pictus D. Don that can be administered for prevention,treatment and optimum glycemic control of diabetes mellitus in humansand mammals. The disclosure provides an extract of Costus pictus D. Don.The disclosure provides a medicinal preparation of Costus pictus D. Don.The disclosure provides a nutritional composition Costus pictus D. Don.The disclosure provides combinations of medicinal preparation andnutritional composition of Costus pictus D. Don.

Methods to control blood glucose concentration by moderateadministration of the disclosed medicinal preparation are disclosed. Inappropriate proportions and with right techniques the medicinalpreparation can be administered to regenerate beta cells of pancreaticislets. An innovative extraction of this innovative preparationadministered therapeutically deploying an innovative manner and methodeffectively controls glycogen level in the liver of mammals and humans.

The disclosure provides an extract of Costus pictus D. Don, whichaddresses the various aspects of diabetes concurrently, namely, controlof blood sugar, regeneration of the beta cells of pancreas resulting inincreased secretion of insulin, reducing insulin resistance, andcorrecting decreased hepatic glycogen synthesis. A key aspect of thepresent invention, thus, is diabetes management. Glycogen control formsone aspect of this management. The disclosure provides a compositionfrom costus which possesses sufficient novelty by favorably modifyingthe different aspects of the disease, simultaneously.

The disclosure teaches an innovative and path breaking technique totreat Diabetes mellitus by obtaining optimum glycemic control in blood.The disclosure teaches a plant extract in the form of pharmaceuticalcompositions and therapeutically (or prophylactically) effective amountof the compositions that is also pharmaceutically acceptable. Thecarrier and composition are produced under good laboratory practicesconditions. The pharmaceutical preparation is ideally selected such thatit suits the mode of administration, in accordance with conventionalpractice.

The preparation, if desired, can also contain minor amounts ofwetting/emulsifying agents or pH buffering agents. In addition, thepreparation can be a dosage form such as a liquid solution, suspension,emulsion, tablet, pill, capsule, sustained release formulation, powder,paste, ointment, sachet, ampoule, injection, infusion or combinationsthereof.

Further, the compositions can be formulated in accordance with methodsthat are well known in the art of pharmaceutical composition and can besuitably adapted for intramuscular/subcutaneous/intravenousadministration to human beings. Generally, the ingredients are suppliedeither separately or mixed together in unit dosage form, for example, asa dry lyophilized powder or water free concentrate in a hermeticallysealed container such as an ampoule or sachet indicating the quantity ofactive compound. Where the composition is to be administered byinfusion, it can be dispensed with an infusion bottle containing sterilepharmaceutical grade water, saline or dextrose/water. Where thecomposition is administered by injection, an ampoule of sterile waterfor injection or saline can be provided so that the ingredients may bemixed just prior to administration. Finally, compositions of theinvention can be formulated as neutral or salt forms.

The amount of the invention's composition, which will be effective inthe treatment of a particular disorder or condition, can be determinedby standard clinical techniques. In addition, in vitro and/or invivo'assays may optionally be employed to help identify optimal dosageranges. The precise dose to be employed in the formulation will alsodepend on the route of administration and the seriousness of the diseaseor disorder. The dose should be decided according to the judgment of thepractitioner taking into consideration each patient's circumstances.

The disclosure provides a method and technique of treatment, preventionand/or management of diabetes wherein a hypoglycemically effectivemedicinal and/or nutritional preparation of Cactus pictus D. Don can beadministered to humans. The disclosure provides an effective treatmentfor controlling glycogen levels in the liver of diabetic mammals andhumans. In an exemplary method the preparation is administered orally.The disclosure teaches a method and technique of administering thepreparation of Costus pictus D. Don in the dosage of 500 mg-2000 mg/dayfor controlling type I and type II diabetics in human beings in the formof capsules, tablets, granules, sachets, powder and such othervariations that may become available in future to fulfill the objectivesof treatment, management or prevention of diabetes.

An ideal dosage includes administration of about 500 mg/day to about2000 mg/day of an extract of Costus pictus D. Don to a mammal in needthereof for treatment, management or prevention of type I or type IIdiabetes. Other dosages include administration of about 50 mg/day toabout 200 mg/day to a mammal in need thereof. In some embodiments, themethod of treatment of diabetes mellitus includes administering a doseof extract of about 50 mg/kg/day to about 200 mg/kg/day to astreptozotocin induced diabetic rat. In some embodiments of the methodof treatment of diabetes mellitus, a dose of extract of ranging fromabout 500 mg/day to about 2000 mg/day is administered to a human. Inexemplary method of treatment of diabetes mellitus, the extract isadministered at a dose ranging from about 500 mg/day to about 2000mg/day to a diabetic patient. In some embodiments of the method ofprevention of diabetes mellitus, a dose of the extract ranging fromabout 500 mg/day to about 2000 mg/day is administered to a human. Insome embodiments of the method of management of diabetes mellitus, adose of the extract of about 500 mg/day to about 2000 mg/day isadministered to a human.

The extract can be administered to a mammal for a method of preventingdiabetes. The extract can be administered to a mammal for a method ofmanagement of diabetes. In an exemplary method the extract isadministered orally. In an exemplary method, the method of management ofdiabetes includes glycemic control. In an exemplary method, the methodof management of diabetes includes regeneration of the beta cells ofpancreas. In an exemplary method, the method of management of diabetesincludes enhancing the capacity of pancreatic beta cells by causingendogeneous insulin producing cells to become more effective insulinproducers. In an exemplary method, the method of management of diabetesincludes enhancing hepatic glycogen synthesis from endogenous glucose.In an exemplary method, the method of management of diabetes includesreduced insulin resistance. In an exemplary method, a hypoglycemicallyactive medicinal preparation is administered to humans for management ofdiabetes. In an exemplary method, a hypoglycemically active medicinalpreparation is administered to humans for treatment of diabetes. Theextract can be administered to a mammal for a method of treatment ofdiabetes. The extract can be administered to a mammal for a method ofinducing hypoglycemia. The extract can be administered to a mammal for amethod of regenerating beta-cells of pancreas. An exemplary method forincreasing the capacity of insulin producing cells (β cells of Islets ofLangerhans) in humans and animals is administering a therapeuticallyeffective dose of a composition of an extract of Costus pictus D. Don.The extract can be administered to a mammal for a method for increasingthe capacity of insulin producing cells of the mammal to become moreeffective insulin producers. The extract can be administered to a mammalfor a method for increasing the capacity of insulin producing cells,whereby damaged beta-cells present in the pancreas regenerate intoinsulin producing cells. The extract can be administered to a mammal fora method of controlling normal liver glycogen in the mammal. Aninnovative method, technique and treatment to regenerate beta cells ofpancreatic islets includes administering an extract of Costus pictus D.Don in the dose of 50-200 mg/kg/day in streptozotocin induced diabeticrats and such other mammals and also extendable to humans in applicabledosages including a dosage of 500-2000 mg/day for humans. The disclosureprovides ecofriendly and value engineered methods for preparing aneffective extract from plant Costus pictus D. Don to obtain optimumglycemic control.

In some embodiments of the method of inducing hypoglycemia, a close ofthe extract of about 500 mg/day to about 2000 mg/day is administered toa human. In some embodiments of the method of regenerating beta-cells ofpancreas, a dose of the extract of about 500 mg/day to about 2000 mg/dayis administered to a human. In some embodiments of the method forincreasing the capacity of insulin producing cells (β cells of Islets ofLangerhans) in a human, a dose of the extract of about 500 mg/day toabout 2000 mg/day is administered to a human. In some embodiments of themethod for increasing the capacity of insulin producing cells in ahuman, whereby damaged beta-cells present in the pancreas regenerateinto insulin producing cells, a dose of the extract of about 500 mg/dayto about 2000 mg/day is administered to a human. In some embodiments ofthe method of controlling normal liver glycogen in the mammal, a dose ofthe extract of about 500 mg/day to about 2000 mg/day is administered toa human.

The disclosure provides methods for long term management of diabetes byavoiding the problems associated with tight control of blood glucoseconcentrations, i.e., hypoglycemia tolerance and seizures, whilesimultaneously avoiding the problems associated with conventionalmoderate control of blood glucose concentrations, i.e., pathologicalcomplications associated with hyperglycemia, such as nephropathy,retinopathy etc.

In some embodiments, the extract can be effectively mixed with otherherbal/nutraceutical/synthetic components without losing itsobjectivity.

The disclosure provides a path breaking method wherein the innovativemedicinal preparation of Costus pictus D. Don is obtained by using lowmolecular weight alcohols, as such or mixed with water, halogenatedhydrocarbons, organic ethers, low molecular weight esters, other organicsolvents and low molecular weight ketones. The disclosure providesmethods for preparing extracts of Costus pictus D. Don. An exemplarymethod of preparing an extract of Costus pictus D. Don includescollecting a starting material of Costus pictus D. Don; treating thestarting material with low molecular weight alcohols, low molecularweight alcohols mixed with water, halogenated hydrocarbons, organicethers, low molecular weight esters, alkalies, other organic solvents,low molecular weight ketones, water or combinations thereof to prepare amixture; and processing the mixture to prepare the extract. The startingmaterial includes a fresh whole plant of Costus pictus D. Don, freshleaves of Costus pictus D. Don, dried leaves of Costus pictus D. Don,and combinations thereof.

Low molecular weight alcohols that can be used in preparation of theextract include methanol, ethanol, isopropanol, n-butanol andcombinations thereof. Halogenated hydrocarbons that can be used forextract preparation include methylene chloride, ethylene dichloride,chloroform, and combinations thereof. Esters that can be used forextract preparation include methyl acetate, ethyl acetate, propylacetate, n-butyl acetate and combinations thereof. Ketones that can beused for extract preparation include acetone, methyl ethyl ketone, andcombinations thereof. Alkanes that can be used for preparation of theextract include pentane, hexane, heptane, isooctane, and combinationsthereof.

An exemplary method of preparing an extract using fresh leaves of Costuspictus D. Don as the starting material is provided in FIG. 1. In FIG. 1,fresh leaves of Costus pictus D. Don are subjected to extraction withmethanol to obtain a mixture. The mixture is filtered to obtain afiltrate and a residue (Residue I). The filtrate is the methanol solublefraction (Fraction I or Fr. I). Fraction I is concentrated and dried toobtain an extract. The extract Fraction I is reextracted with hexane toobtain a mixture. The hexane containing mixture is filtered to obtain afiltrate, which is a hexane soluble fraction (Fraction II or Fr. II),and a residue (Residue II). Next Residue II is further extracted withethyl acetate to form a mixture. The ethyl acetate containing mixture isfiltered resulting in a filtrate, which is an ethyl acetate solublefraction (Fraction III or Fr. III), and a residue (Residue III). TheResidue III is extracted with butanol to prepare a mixture. The butanolcontaining mixture is filtered resulting in a filtrate, which is abutanol soluble fraction (Fraction IV or Fr. IV), and a residuefraction, namely Fraction V. Fractions II, III, IV and IV areconcentrated and dried to obtain extracts. The dried extracts aredissolved in water or other suitable solvents to test for hypoglycemicactivity in mammals.

An exemplary method of preparing an extract of Costus pictus D. Donincludes collecting a starting material comprising Costus pictus D. Don;refluxing the starting material with a first low molecular weightalcohol to prepare a mixture I; filtering the mixture I to prepare afiltrate I and a residue I; concentrating the filtrate I to prepare aconcentrate I; and drying the concentrate I to prepare a fraction I,wherein the starting material includes a fresh whole plant of Costuspictus D. Don, fresh leaves of Costus pictus D. Don, dried leaves ofCostus pictus D. Don, or combinations thereof, and wherein the fractionI is designated as the extract.

An exemplary method of preparing an extract includes treating fraction Iwith an alkane to prepare a mixture II; filtering the mixture II toprepare a filtrate II and a residue II; concentrating the filtrate II toprepare a concentrate II; and drying the concentrate II to form afraction II, wherein the fraction II is designated as the extract.

An exemplary method of preparing an extract includes treating residue IIwith a low molecular weight ester to prepare a mixture III; filteringthe mixture III to prepare a filtrate III and a residue III;concentrating the filtrate III to prepare a concentrate III; and dryingthe concentrate III to form a fraction III, wherein the fraction III isdesignated as the extract.

An exemplary method of preparing an extract includes treating residueIII with a second low molecular weight alcohol to prepare a mixture IV;filtering the mixture IV to prepare a filtrate IV and a residue(fraction V), wherein the residue (fraction V) is designated as theextract. An exemplary method of preparing an extract includesconcentrating filtrate IV to prepare a concentrate IV; and drying theconcentrate IV to form a fraction IV, wherein the fraction IV isdesignated as the extract.

In an exemplary method of preparing the extract the first low molecularweight alcohol is mixed with water.

An exemplary method of preparing an extract includes collecting freshleaves of Costus pictus D. Don, cleaning the fresh leaves to preparecleaned leaves, drying the cleaned leaves to prepare dried leaves,powdering the dried leaves to prepare a dried powder. In an exemplarymethod of preparing the extract, the dried powder is refluxed with thefirst low molecular weight alcohol to prepare an extract.

First low molecular weight alcohols that can be used for preparing anextract include methanol, ethanol, isopropanol, and combinationsthereof.

Second low molecular weight alcohols that can be used for preparing anextract include methanol, ethanol, propanol, n-butanol, and combinationsthereof.

Low molecular weight esters that can be used for preparing an extractinclude methyl acetate, ethyl acetate, propyl acetate, n-butyl acetateand combinations thereof.

Alkanes that can be used for preparing an extract include pentane,hexane, heptane, isooctane, and combinations thereof.

In an exemplary method of preparing the extract, the starting materialof Costus pictus D. Don is a fresh whole plant of Costus pictus D. Don.In an exemplary method of preparing the extract, the starting materialis fresh leaves of Costus pictus D. Don. In an exemplary method ofpreparing the extract, the starting material is dried leaves of Costuspictus D. Don. In an exemplary method of preparing the extract, thestarting material is a combination of two or more of fresh plant ofCostus pictus D. Don, fresh leaves of Costus pictus D. Don and driedleaves of Costus pictus D. Don.

INDUSTRIAL APPLICATION OF INVENTION

Accordingly it is an object of this invention to provide an improvedmethod of preparing a substance for treating diabetes mellitus.

Another object of the present invention is to provide methods oftreatment for controlling/preventing type I and type II diabeticpatients without side effects.

A feature of present invention is to regenerate pancreatic beta cellsand obtain optimum glycemic control in human beings.

It will be readily understood by the skilled artisan that numerousalterations may be made to the examples and instructions given hereinincluding the generation of different DP IV inhibitors and alternatetherapeutic compositions without departing from either the spirit orscope of the present invention. These and other objects and features ofpresent invention will be made apparent from the following examples. Thefollowing examples as described are not intended to be construed aslimiting the scope of the present invention.

EXAMPLES Example 1 Preparation of Dry Powder

Fresh leaves of Costus pictus were collected, cleaned and dried undershade. The dried leaves were powdered and kept as dried powder.

Example 2 Preparation of Extract

An extract of Costus pictus D. Don was prepared according to theprocedure shown in FIG. 1. Fresh leaves of Costus pictus D. Don wasrefluxed with 90% methanol for four hours. After refluxing the mixturewas filtered and filtrate was concentrated and dried (fraction I or Fr.I). This extract was fractionated with different solvents like hexane,ethyl acetate and butanol. The fraction II or Fr. II (hexane part),fraction III or Fr. III (ethyl acetate part), fraction IV or Fr. IV(butanol part) and fraction V or Fr. V (residue) were concentrated anddried under vacuum. The hexane soluble fraction (II) was discarded. Theresidue was extracted with ethylacetate. The fractions I, III, IV and Vwere screened for hypoglycemic activity.

Example 3

Activity of different extracts were tested using Sprague dawley rats ofeither sex aged 3 months weighing 180-200 g. After 3-4 hrs of fastingthe rats were divided into 7 groups of 6 in each, one group kept ascontrol and 4 groups were given different fractions (Fraction I,Fraction III, Fraction IV and Fraction V) of extract in the dose of 100mg/kg. Sixth and seventh groups were given standard drugs glybenclamide(1 mg/kg) and metformin (150 mg/kg) respectively.

As seen in Table 2 and the graph in FIG. 2, results of the study showedthat administration of fraction III (ethyl acetate part, shown inFIG. 1) and fraction V (residue, shown in FIG. 1) in the dose of 100mg/kg significantly (p<0.05) decreased the blood glucose level afterfive hours. In the case of standard drug glybenclamide there wassignificant (p<0.05) decrease in blood glucose level in the first hourand effect was more pronounced in the third and fifth hour. Metforminhowever produced significant (p<0.05) decrease in blood glucose level inthe fifth hour. (See Table II, graph I) The extract/drug wasadministered orally (single dose) after 4 hours of fasting. Fraction III(ethyl acetate fraction) and fraction V (residue) showed significantdecrease (p<0.05) in blood glucose level after 5 hours after extractfeeding. Standard drug glybenclamide produced significant (p<0.05)decrease in blood glucose level in the first hour itself, whilemetformin produced significant decrease (p<0.05) in the fifth hour. InFIG. 2, the graph provides blood glucose level in normal rats in mg/dlmeasured over a duration of 5 hours. The normal rats were fed withdifferent fractions of the extract after four hours of fasting. Thegraph shows that administration of Fraction III and Fraction V produceda significant decrease in blood glucose level after 5 hours.

TABLE 2 Effect of different fractions of extract of Costus pictus D. Donon blood sugar level (mg/dl) in normal rats. Group Dosage/kg Fasting 1hr 3 hr 5 hr Control 1 ml 75.3 ± 5.4 65.6 ± 4.2 64.0 ± 4.3  62.0 ± 3.3distilled water Fraction I 100 mg 73.5 ± 0.6 76.8 ± 2.8 68.5 ± 4.5  69.5± 4.6 Fraction III 100 mg 78.8 ± 3.5 68.1 + 4.8 68.0 ± 5.3 *63.8 ± 5.6Fraction IV 100 mg 67.1 ± 2.2 62.5 ± 3.1 63.3 ± 2.7  60.5 ± 1.9 FractionV 100 mg 70.3 ± 4.3 67.3 ± 1.0 62.0 ± 2.0 *57.0 ± 2.3 Glybenclamide  1mg 72.6 ± 8.6 *58.8 ± 9.8  *43.3 ± 8.2  *43.2 ± 5.8 Metformin 150 mg76.6 ± 4.1 70.6 ± 3.0 68.3 ± 2.6 *61.5 ± 3.0

The most active fraction, i.e., ethyl acetate fraction (fraction III)was taken for further animal experiments and human trials.

Example 4

Glucose tolerance study of extract of Costus pictus D. Don was conductedin normal Sprague dawley rats. After acclimatization period, rats weredivided into five groups—one group (Group A) kept as control andremaining four groups (Group B, Group C, Group D and Group E) asexperimental groups. Fasting blood glucose was noted and experimentalanimals was administered the extract (ethyl acetate fraction) orally inthe dose of 50 mg (Group B), 100 mg (Group C), 200 mg (Group D), and 1g/kg (Group E). Glucose was administered orally in the dose of 1 g/kgfor all animals one hour after extract administration. Blood sugar wastested at an interval of half hour, 1 hour, 2 hour, 3 hour and 5 hour bythe method of Asatoor and King.

Results of glucose tolerance test using different doses of the abovesaid extract showed that doses of 100 mg/kg and 200 mg/kg inhibited theinitial peak rise in blood glucose and produced significant (p<0.05)decrease in blood glucose level within 3 hrs after glucoseadministration (Table 3, FIG. 3). Oral administration of extract in thedose of 100 mg and 200 mg/kg produced significant (p<0.05) decrease inthe 3rd and 5th hour in normal rats. The dosages 50 mg/kg and 1 g/kg didnot produce any significant effect. The results of the glucose tolerancetest in normal rats are depicted graphically in FIG. 3. FIG. 3 showsthat administration of extract of Costus pictus D. Don in the dose of100 mg/kg and 200 mg/kg produced significant decrease in blood glucosewithin 3 hrs after glucose loading.

TABLE 3 Effect of extract of Costus pictus D. Don on GTT (glucosetolerance test) in normal rats. Glucose administration after 1 hr afterextract administration Fasting ½ hr 1 hr 2 hr 3 hr 5 hr group dosage(mg/dl) (mg/dl) (mg/dl) (mg/dl) (mg/dl) (mg/dl) Group A 1 ml distilled55.4 ± 5.4  79.0 ± 6.05  71 ± 2.9 58.0 ± 3.8 50.3 ± 4.1 52.2 ± 4.8 waterGroup B  50 mg/kg 53.7 ± 5.2 66.2 ± 3.4 59.5 ± 2.4 55.8 ± 3.3 53.5 ± 0.949.8 ± 1.2 Group C 100 mg/kg 70.5 ± 3.6 66.5 ± 1.9 60.8 ± 2.4 61.5 ± 1.1*57.0 ± 3.9  *47.7 ± 4.3  Group D 200 mg/kg 62.8 ± 3.5 63.7 ± 2.6 60.3 ±2.4 57.0 ± 1.6 *52.2 ± 1.7  *47.8 ± 1.2  Group E  1 g/kg 58.8 ± 3.0 89.3 ± 10.2 76.3 ± 7.8 62.2 ± 5.7 61.0 ± 7.0 54.8 ± 1.2

Example 5

Glucose tolerance study of extract of Costus pictus D. Don was conductedin experimental diabetic rats. Diabetes was induced by injectingStreptozotocin (50 mg/kg—intraperitoneally) and they were divided intotwo groups. One group (Group B, “Str+ ext”) was given the above extractin the dose of 100 mg/kg and second group (Group C, “Str”) maintained asuntreated diabetic control. Another group was administered distilledwater served as the normal control group (Group A). After 35 days oforal treatment of extract these animals were subjected to glucosetolerance test along with normal control group (Group A).

Glucose tolerance test (GTT) results provided in Table 4 and graph inFIG. 4 show that in untreated diabetic control animals, blood glucoseincreased to a significant (p<0.05) level within half an hour afterglucose administration and remained significantly high throughout theexperimental period. In the extract treated diabetic rats, there wasinhibition of initial hyperglycemia and blood glucose returned tobaseline level within three hours after glucose administration similarto normal control rats. Extract administration prevented initialhyperglycemia and blood sugar level became normal within three hours,while in untreated diabetic control blood sugar level increased to asignificant level and remained high throughout the experimental period.In FIG. 4, the graph shows glucose tolerance test in diabetic rats,wherein glucose concentration in blood in mg/dl was measured over aduration of 5 hrs. The results show that oral administration of 100mg/kg of Costus pictus D. Don prevented hyperglycemia in diabetic rats(Group B). Whereas after glucose loading, the blood glucose increasedsignificantly throughout the experimental period in the untreateddiabetic control rats (Group C).

TABLE 4 Effect of extract of Costus pictus D. Don on GTT in Diabetic andTreated Rats Glucose administration (1 g/kg) 1 hr after extractadministration Fasting ½ hr 1 hr 2 hr 3 hr 5 hr Groups (mg/dl) (mg/dl)(mg/dl) (mg/dl) (mg/dl) (mg/dl) Group A 54.4 ± 3.1 75.5 ± 4.8 73.3 ± 3.165.5 ± 3.1 63.7 ± 1.7 62.2 ± 2.1 (control) Group B 51.6 ± 4.5 55.2 ± 5.963.2 ± 8.1 57.0 ± 5.5 50.4 ± 2.1 50.4 ± 4.2 (Str + ext) Group C 104.2 ±20.7 *164.0 ± 52.2  *146.0 ± 41.6  *129.4 ± 41.1  *135.0 ± 46.6  *144.8± 44.9  (Str)

Example 6

Hypoglycemic action of extract of Costus pictus D. Don were tested inStreptozotocin (50 mg/kg) induced diabetic rats of Sprague dawley strainof either sex. They were randomly assigned to three groups—(A) controlgroup, (B) extract treated group and (C) untreated diabetic control.Diabetes was induced in Groups B and C by an injection of streptozotocinfor 10 days.

Blood was collected from the caudal vein under light ether anaesthesiaafter an overnight fasting. Blood glucose was estimated (Asatoor & King)before starting the experiment and repeated every 7 days for 5 weeks.Liver glycogen was estimated in all animals after sacrificing theanimals. Pancreas from the animals were processed for histologicalstudies. Body weight of all animals were noted before and every 10 daysduring administration of extract.

Results in Table 5 and graph in FIG. 5 show that blood glucose levelremained same throughout the experimental period in control rats. Therewas a significant increase in blood sugar level in group B and C whichwere given streptozotocin. Administration of extract (100 mg/kg orally)produced decrease in blood glucose level within three days itself, butnot to a significant level. Seven days treatment produced significantdecrease (p<0.05) in blood glucose level, and twenty eight daystreatment brought the blood glucose to baseline level. Further treatmentdid not produce hypoglycemia. Blood glucose level remained inhyperglycemic state in group C (untreated diabetic rats), throughout theexperimental period. Table 5 and graph in FIG. 5 show that extractadministration caused a significant decrease (p<0.05) in blood glucoselevel by 7 days treatment, glucose became normal within 28 daystreatment and further treatment did not produce hypoglycemia. Inuntreated diabetic rats blood sugar level remained high compared to thetreatment group throughout the study period. The graph in FIG. 5 showsthat oral administration of extract of Costus pictus D. Don (100 mg/kg)significantly decreased glucose level in streptozotocin induced diabeticrats (group B), whereas in untreated diabetic control (group C) bloodglucose remained significantly higher throughout the experimentalperiod.

TABLE 5 Effect of extract of Costus pictus D. Don on streptozotocin (50mg/kg) induced diabetic rats 10 days after Treatment periodstreptozotocin 3 7 14 21 28 35 Groups Fasting injection days days daysdays days days Group A 72.1 ± 5.3 70.0 ± 3.4 72.0 ± 3.0 73.1 ± 4.2 72.7± 3.1 73.0 ± 4.1 73.7 ± 4.1 72.2 ± 3.8 (control) Group B 72.0 ± 3.8*223.0 ± 20.9  219.0 ± 20.1 *179.1 ± 20.6  *131.0 ± 18.2  *85.0 ± 7.4 *73.0 ± 6.0  *73.0 ± 4.0  (Str + ext) Group C 70.0 ± 4.8 249.0 ± 26.1246.0 ± 26.6 243.0 ± 26.7 226.9 ± 25.4 217.0 ± 25.0 207.0 ± 21.0 141.0 ±19.4 Str

Liver glycogen level in extract (100 mg/kg) treated groups was same asin normal control rats. But there was significant decrease (p<0.05) inglycogen level in untreated diabetic control rats. See Table 6 and graphin FIG. 6. Low liver glycogen in group C (untreated diabetic rats)showed decreased conversion of blood glucose to liver glycogen in thediabetic rats due to absence of insulin. The extract seems to increasethe conversion of glucose to glycogen in the liver of diabetic rats.Table 6 shows that liver glycogen level in untreated diabetic controlrats significantly (p<0.05) decreased, while in the treatment groupglycogen level was brought back to normal as in control group. The graphin FIG. 6 shows significantly (p<0.05) lower liver glycogen level inuntreated diabetic control (group C) than normal (group A) and treatmentgroup (group B).

TABLE 6 Effect of extract of Costus pictus D. Don on liver glycogen atthe end of treatment period. Group A Group B Group C (control) (Str +ext) (Str) 30.5 ± 0.3 32.3 ± 1.1 *6.0 ± 1.4

Histopathological studies showed decreased number of islets ofLangerhans and decreased numbers of beta cells per islets in theuntreated diabetic control compared to normal or extract treated groups.In diabetic rats varying degrees of cellular damage was seen in theislets. There was decrease in population of cells with nuclear pyknosis,karyorerrhsis and karyolysis with degranulation and cytolysis. Intreatment group the degree of cellular damage was lesser andregeneration was observed. Compared to normal rats, there was 75%decrease in number of cells in untreated diabetic control, while intreatment group there was only 29% decrease. Table 7 shows thattreatment of diabetic rats with extract showed regeneration ofpancreatic beta cells in Islets, while in untreated diabetic controlnumber of beta cells was less.

TABLE 7 Effect of extract of Costus pictus D. Don treatment on number ofbeta cells per pancreatic islets in experimental rats Effect oftreatment on No. of beta pancreatic cells compared Group cells/islets tonormal (percentage) Normal 139 ± 17.4 Treatment group  98 ± 21.5 29%Untreated diabetic control 34 ± 8.5 75%

Table 8 shows that administration of extract in the dose of 100 mg/kginhibited the weight loss in diabetic rats (group B) even by 10^(th) dayand weight was brought back to baseline level by 20^(th) day oftreatment. Rats administered with distilled water served as control(Group A) did not exhibit any weight loss. In untreated diabetic control(Group C) the body weight was significantly lower than normal control.The graph in FIG. 7 shows that administration of extract of Costuspictus D. Don brought the body weight to normal level within 20 daystreatment in group B, while body weight remained significantly lower ingroup C.

TABLE 8 Effect of extract of Costus pictus D. Don on body weight (grams)in different groups of rats 10 days Treatment period Groups Baselineafter Str 10 days 20 days 30 days 35 days Group A 318.6 ± 10.5   323 ±9.5   332 ± 11.1  *337 ± 11.1 *342.5 ± 10.7 *347.5 ± 9.8  Group B 305.3± 7.0  *272.0 ± 6.6  296.5 ± 10.8 309.8 ± 8.0 313.2 ± 8.9 305.0 ± 7.5Group C 314.8 ± 11.6 *285.5. ± 8.6  *272.6 ± 10.1 *255.5 ± 9.2  *233.8 ±9.8  *222.8 ± 11.2

Example 7

Acute toxicity study of extract of Costus pictus D. Don were tested inSwiss albino mice of both sexes. The mice divided into three groups ofsix in each. Group A was kept as control group and remaining 2 groups (Band C) were used for experiment. Group A mice were given distilled water(0.25 ml/25 g body weight) and groups B and C were given 0.25 ml extractper 25 g mice in the dosage of 100 mg and 200 mg/Kg body weight orallyand observed continuously for first six hours. Mortality was recordedfor 15 days. (See Ghosh, 1970)

The extract was non-toxic even up to 200 mg/kg given orally to albinomice. All animals behaved normally. No neurological or behaviouraleffects were noted. No mortality was recorded in 15 days. This studyshowed that oral feeding of extract of Costus pictus D. Don even up to200 mg/kg did not produce any acute toxic effects in mice.

Example 8

Effect of extract of Costus pictus D. Don was tested in a normal healthyfemale subject (52 yrs). After estimating fasting blood glucose, 500 mgof extract (I capsule) was given to normal subject, before breakfast.Blood sugar was estimated after 2 hr, 5 hr and subject was allowed totake normal diet. Fasting blood glucose was estimated after 24 hr also.(See Table 9 and graph in FIG. 8)

Results provided in Table 9 and graph in FIG. 8 showed that capsule didnot have any significant effect on normal subject. Table 9 shows thatadministration of 500 mg of extract (single dose) did not produce anysignificant effect on blood glucose level in normal subject.

TABLE 9 Effect of 500 mg extract (1 capsule - single dose) of Costuspictus D. Don on blood glucose (mg/dl) level in normal Human beingCapsule + normal breakfast diet subject Fasting 2 hr 5 hr 24 hr normal(1) 88 95 88 95

Example 9

Effect of extract of Costus pictus D. Don was tested in diabeticpatients (45-52 yrs). After estimating fasting blood glucose, 500 mg ofextract (I capsule) was given to patient, before breakfast. Blood sugarwas estimated after 2 hr, 5 hr and patients were allowed to take normaldiet. Fasting blood glucose was estimated after 24 hr also.

Table 10 shows that administration of 500 mg extract (single dose)produced significant effect on blood glucose level in diabetic patient.Result showed significant lowering of blood glucose from the 2nd houronwards, which reached maximum on 5th hour period in diabetic subjects.Results are also provided in the graph in FIG. 8. FIG. 8 shows theeffect of 500 mg of extract of Costus pictus D. Don (single dose) onnormal (group A) and diabetic (group B) human beings.

TABLE 10 Effect of 500 mg extract (1 capsule - single dose) of Costuspictus D. Don on blood glucose (mg/dl) level in diabetic patientsCapsule + normal breakfast diet subject fasting 2 hr 5 hr 24 hrDiabetics(5) 130.7 ± 1 144.3 ± 26.1 98.7 ± 5.3 100 ± 12.5

Other modifications and variations to the invention will be apparent tothose skilled in the art from the foregoing disclosure and teachings.Thus, while only certain embodiments of the invention have beenspecifically described herein, it will be apparent that numerousmodifications may be made thereto without departing from the spirit andscope of the invention.

1. A method of preparing an extract of Costus pictus D. Don comprising:collecting a starting material comprising Costus pictus D. Don;refluxing the starting material with a low molecular weight alcohol toprepare a mixture I; filtering the mixture I to prepare a filtrate I anda residue I; concentrating the filtrate I to prepare a concentrate I;and drying the concentrate I to prepare an extract (fraction I).
 2. Themethod of claim 1 further comprising: treating the extract (fraction I)with an alkane to prepare a mixture II; filtering the mixture II toprepare a filtrate II and a residue II; concentrating the filtrate II toprepare a concentrate II; and drying the concentrate II to prepare anextract (fraction II).
 3. The method of claim 2 further comprising:treating the residue LI with a low molecular weight ester to prepare amixture III; filtering the mixture III to prepare a filtrate III and aresidue III; concentrating the filtrate III to prepare a concentrateIII; and drying the concentrate III to prepare an extract (fractionIII).
 4. The method of claim 1, wherein the low molecular weight alcoholis mixed with water to perform the refluxing.
 5. The method of claim 1,wherein the low molecular weight alcohol is selected from a groupconsisting of methanol, ethanol, isopropanol, and combinations thereof.6. The method of claim 1, wherein the low molecular weight alcohol is90% methanol.
 7. The method of claim 1, wherein said starting materialis selected from the group consisting of a fresh whole plant of Costuspictus D. Don, fresh leaves of Costus pictus D. Don, dried leaves ofCostus pictus D. Don, or combinations thereof.
 8. The method of claim 2,wherein the alkane is selected from the group consisting of pentane,hexane, heptane, isooctane, and combinations thereof.
 9. The method ofclaim 3, wherein the low molecular weight ester is selected from thegroup consisting of methyl acetate, ethyl acetate, propyl acetate,n-butyl acetate and combinations thereof.
 10. A method of preparing anextract of Costus pictus D. Don, the method comprising: collecting freshleaves of Costus pictus D. Don; cleaning the fresh leaves to preparecleaned leaves; drying the cleaned leaves to prepare dried leaves;powdering the dried leaves to prepare a dried powder, and, refluxing thedried powder with a low molecular weight alcohol to prepare a mixture I;filtering the mixture I to prepare a filtrate I and a residue I;concentrating the filtrate I to prepare a concentrate I; and drying theconcentrate I to prepare an extract (fraction I).
 11. The method ofclaim 10 further comprising: treating the extract (fraction I) with analkane to prepare a mixture II; filtering the mixture II to prepare afiltrate II and a residue II; concentrating the filtrate II to prepare aconcentrate II; and drying the concentrate II to prepare an extract(fraction II).
 12. The method of claim 10 further comprising: treatingthe residue II with a low molecular weight ester to prepare a mixtureIII; filtering the mixture III to prepare a filtrate III and a residueIII; concentrating the filtrate III to prepare a concentrate III; anddrying the concentrate III to prepare an extract (fraction III).
 13. Themethod of claim 10, wherein said the low molecular weight alcohol ismixed with water to perform the refluxing.
 14. The method of claim 10,wherein the low molecular weight alcohol is selected from a groupconsisting of methanol, ethanol, isopropanol, and combinations thereof.15. The method of claim 10, wherein the first low molecular weightalcohol is 90% methanol.
 16. The method of claim 11, wherein the alkaneis selected from the group consisting of pentane, hexane, heptane,isooctane, and combinations thereof.
 17. The method of claim 12, whereinthe low molecular weight ester is selected from the group consisting ofmethyl acetate, ethyl acetate, propyl acetate, n-butyl acetate andcombinations thereof.